Comparison of granulosa-cell derived factors in response to FSH treatment in WT and AHRKO follicles. Early antral follicles from pre-pubertal WT and AHRKO ovaries were isolated and cultured in supplemented media in the presence of 0–15 IU/mL FSH for 7 days. At the end of culture, follicles were pooled per treatment group and genotype, subjected to extraction of RNA and then processed for qPCR analysis of Fshr (A) and Cyp19a1 (B). Each gene was expressed as a mean relative expression ratio normalized to Actb as a loading control. Media samples from individual follicles were obtained from cultures and subjected to ELISAs to measure levels of estradiol (C). Each bar represents the mean ± SEM from three separate cultures per genotype at each selected treatment and from 10–18 follicles per treatment. The letter “a” above the bars indicates significant difference (p ≤ 0.05) from WT follicles within the same FSH treatment, the letter “b” above the bars indicates significant difference (p ≤ 0.05) from 0 IU/mL FSH in the same genotype and letter “c” above the bar indicates significant difference (p ≤ 0.05) from WT follicles treated with 5 IU/mL FSH, using one-way ANOVA followed by Tukey’s test as a post hoc test.